Advancing biological and clinical applications of genomic Minimal Residual Disease detection in AML

推进基因组微小残留病检测在 AML 中的生物学和临床应用

基本信息

批准号：
9763499
负责人：
Sami Nimer Malek
金额：
$ 41.69万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-01 至 2023-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9763499
关键词：
Acute Myelocytic Leukemia Adult Affect Age Alleles Allogeneic Bone Marrow Transplantation Biological Biological Assay Biological Markers Blast Cell Bone Marrow Bone Marrow Cells Cell Separation Cells Cessation of life Chemotherapy-Oncologic Procedure Clinical Clinical Management Complement Consolidation Therapy Cytogenetics Data Decision Making Detection of Minimal Residual Disease Development Diagnosis Disease remission Foundations Future Gene Frequency Gene Mutation Genes Genomics Hematopoiesis In complete remission Individual Maintenance Maintenance Therapy Marrow Measurement Measures Modeling Monitor Mus Mutate Mutation Mutation Analysis Outcome Patients Phenotype Preleukemia Prognostic Factor Publishing Recurrent disease Relapse Residual Neoplasm Residual Tumors Residual state Risk Sampling Source Techniques Therapeutic Therapeutic Agents Time Tumor Debulking Validation Xenograft procedure age related base chemotherapy clinical application cohort digital disorder risk genetic variant improved insight interest leukemia leukemic stem cell mutant novel novel therapeutic intervention novel therapeutics parkin gene/protein predict clinical outcome prognostic prognostic assays prognostic value single-cell RNA sequencing survival outcome transcriptome transcriptome sequencing

项目摘要

ABSTRACT Acute myelogenous leukemia (AML) affects more than 20,000 adult patients in the US per year causing greater than 11,000 untimely deaths. Standard AML therapy comprises chemotherapy induction to achieve leukemia debulking, followed by a few cycles of consolidation chemotherapy alone and/or followed by allogeneic bone marrow transplantation. This general therapeutic approach to AML is risk-adapted relying principally on well-established prognostic factors, including the type of AML, age, gene mutations and cytogenetic results. Despite best efforts, AML remains incurable in the majority of afflicted patients. A principle barrier to AML cure is disease relapse despite achieving a clinical complete remission (CR) following standard chemotherapy regimens. Of substantial interest therefore, is the identification and accurate measurement of residual AML that persists during remission in AML patients, as such residual disease is likely the source of relapse. Several important questions about AML relapse are only in early stages of satisfactory answers, including i) how to best measure minimal residual disease (MRD) in AML, ii) whether all MRD is indeed caused by residual disease or rather as suggested by recent data comprises a mixture of leukemia, pre- leukemia and age-related clonal hematopoiesis, iii) what constitutes the cellular source of relapse in AML, iv) if such relapse-causing cells can be better targeted using novel therapeutic approaches, and v) under what circumstances is MRD prognostic and useful for MRD risk-adapted AML clinical management. Given lack of well-suited techniques to reliably answer the relevant questions detailed above, we have optimized droplet digital PCR (ddPCR), a novel ultra-high sensitivity assay for detecting genomic MRD in AML. In a detailed recently published study (Parkin et al, JCI 2017) we found that AML frequently relapses from rare cells residing in remission marrows that based on mutation analysis resemble AML blast cells detected at diagnosis. We demonstrated the feasibility for detecting AML-associated gene mutations at allele frequencies as low as 0.002% and have also provided important novel prognostic insights. In this application, we are proposing complete characterization of mutational MRD and aberrant cellular clusters in AML remission bone marrows using a combination of sophisticated cell sorting and single cell transcriptome analyses complemented with mouse xenografting and ex vivo colony forming assays. Anticipated findings will improve the functional characterization of cells that carry AML-associated gene mutations in an attempt to identify and better characterize the source(s) of AML relapse. Using AML samples from two clinical validation cohorts, we will define the prognostic utility of ddPCR-based MRD assessments. Overall, data will lay the foundation for future real-time genomic MRD-guided clinical AML trials aiming at monitoring and improving consolidation and maintenance therapy and ultimately survival outcome in AML.

抽象的急性骨髓性白血病（AML）每年影响20,000多名成年患者，导致更大超过11,000人过早死亡。标准AML疗法包括化学疗法诱导以实现白血病降临，然后是几个单独的巩固化学疗法和/或接和/或同种异体骨髓移植。这种对AML的一般治疗方法是适应风险的主要是基于公认的预后因素，包括AML的类型，年龄，基因突变和细胞遗传学结果。尽管竭尽全力，但大多数患者中的AML仍然无法治愈。一个 AML治愈的主要障碍是疾病复发，尽管在标准化疗方案。因此，具有实质性的识别和准确是在AML患者缓解过程中持续存在的残留AML的测量，因为这种残留疾病很可能复发的来源。关于AML复发的几个重要问题仅在令人满意的早期阶段答案，包括i）如何最好地测量AML中最小残留疾病（MRD），ii）确实是由残留疾病引起的，或者是最近数据所暗示的，包括白血病的混合物白血病和与年龄相关的克隆造血作用，iii）构成AML复发的构成的原因，iv），如果使用新型的治疗方法可以更好地针对这种引起复发的细胞，v）情况是MRD预后的，对于适应MRD风险的AML临床管理有用。鉴于缺乏适合可靠回答上面详述的相关问题的技术，我们已经优化的液滴数字PCR（DDPCR），这是一种用于检测基因组MRD的新型超高灵敏度测定 AML。在一项详细的最近发表的研究（Parkin等，JCI 2017）中，我们发现AML经常复发从居住在缓解骨髓中的稀有细胞中，基于突变分析类似于AML爆炸细胞在诊断时检测到。我们证明了在等位基因检测AML相关基因突变的可行性频率低至0.002％，还提供了重要的新型预后见解。在此应用中，我们提出了突变MRD和异常细胞的完整表征 AML缓解骨髓中的簇使用复杂的细胞分选和单细胞的组合转录组分析与小鼠异种法和体内菌落形成测定法相辅相成。预期发现将改善携带与AML相关基因突变的细胞的功能表征尝试识别和更好地表征AML复发的来源。使用来自两个临床的AML样品验证队列，我们将定义基于DDPCR的MRD评估的预后效用。总体而言，数据将为未来实时基因组引导的临床AML试验奠定了基础，旨在监视和改善AML的合并和维持疗法，并最终在AML中生存结果。